LOCUS       V021208                 7994 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021208
VERSION     V021208
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..2641
                     /label="TRIM32(NM_012210)"
                     /note="TRIM32(NM_012210)"
                     /gene="TRIM32"
     CDS             2681..2746
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2770..3358
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3424..3472
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3674..4102
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4116..4445
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4512..5306
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5482..5603
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5652..5668)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5676..5692
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5700..5730)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5745..5766
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6054..6642)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6813..7673)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7674..7778)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggctgc agcagcagct tctcacctga acctggatgc
      721 cctccgggaa gtgctagaat gccccatctg catggagtcc ttcacagaag agcagctgcg
      781 tcccaagctt ctgcactgtg gccataccat ctgccgccag tgcctggaga agctattggc
      841 cagtagcatc aatggtgtcc gctgtccctt ttgcagcaag attacccgca taaccagctt
      901 gacccagctg acagacaatc tgacagtgct aaagatcatt gatacagctg ggctcagcga
      961 ggctgtgggg ctgctcatgt gtcggtcctg tgggcggcgt ctgccccggc aattctgccg
     1021 gagctgtggt ttggtgttat gtgagccctg ccgggaggca gaccatcagc ctcctggcca
     1081 ctgtacactc cctgtcaaag aagcagctga ggagcggcgt cgggactttg gagagaagtt
     1141 aactcgtctg cgggaactta tgggggagct gcagcggcgg aaggcagcct tggaaggtgt
     1201 ctccaaggac cttcaggcaa ggtataaagc agttctccag gagtatgggc atgaggagcg
     1261 cagggtccag gatgagctgg ctcgctctcg gaagttcttc acaggctctt tggctgaagt
     1321 tgagaagtcc aatagtcaag tggtagagga gcagagttac ctgcttaaca ttgcagaggt
     1381 gcaggctgtg tctcgctgtg actacttcct ggccaagatc aagcaggcag atgtagcact
     1441 actggaggag acagctgatg aggaggagcc agagctcact gccagcttgc ctcgggagct
     1501 caccctgcaa gatgtggagc tccttaaggt aggtcatgtt ggccccctcc aaattggaca
     1561 agctgttaag aagccccgga cagttaacgt ggaagattcc tgggccatgg aggccacagc
     1621 gtctgctgcc tctacctctg ttacttttag agagatggac atgagcccgg aggaagtggt
     1681 tgccagccct agggcctcac ctgctaaaca gcggggtcct gaggcagcct ccaatatcca
     1741 gcagtgcctc tttctcaaga agatgggggc caaaggcagc actccaggaa tgttcaatct
     1801 tccagtcagt ctctacgtga ccagtcaagg tgaagtacta gtcgctgacc gtggtaacta
     1861 tcgtatacaa gtctttaccc gcaaaggctt tttgaaggaa atccgccgca gccccagtgg
     1921 cattgatagc tttgtgctaa gcttccttgg ggcagatcta cccaacctca ctcctctctc
     1981 agtggcaatg aactgccagg ggctgattgg tgtgactgac agctatgata actccctcaa
     2041 ggtatatacc ttggatggcc actgcgtggc ctgtcacagg agccagctga gcaaaccatg
     2101 gggtatcaca gccttgccat ctggccagtt tgtagtaacc gatgtggaag gtggaaagct
     2161 ttggtgtttc acagttgatc gaggatcagg ggtggtcaaa tacagctgcc tatgtagtgc
     2221 tgtgcggccc aaatttgtca cctgtgatgc tgagggcacc gtctacttca cccagggctt
     2281 aggcctcaat ctggagaatc ggcagaatga gcaccacctg gagggtggct tttccattgg
     2341 ctctgtaggc cctgatgggc agctgggtcg ccagattagc cacttcttct cggagaatga
     2401 ggatttccgc tgcattgctg gcatgtgtgt ggatgctcgt ggtgatctca tcgtggctga
     2461 cagtagtcgc aaggaaattc tccattttcc taagggtggg ggctatagtg tccttattcg
     2521 agagggactt acctgtccgg tgggcatagc cctaactcct aaggggcagc tgctggtctt
     2581 ggactgttgg gatcattgca tcaagatcta cagctaccat ctgagaagat attccacccc
     2641 agaattctgc agatatccag cacagtggcg gccgctcgag gactacaaag accatgacgg
     2701 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     2761 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2821 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2881 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2941 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3001 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3061 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3121 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     3181 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3241 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3301 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     3361 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     3421 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     3481 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     3541 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3601 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     3661 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3721 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3781 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     3841 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3901 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3961 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4021 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     4081 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4141 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4201 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4261 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4321 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4381 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4441 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4501 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4561 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4621 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4681 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4741 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4801 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4861 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4921 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4981 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5041 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5101 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5161 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5221 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5281 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     5341 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5401 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5461 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5521 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5581 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5641 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5701 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5761 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5821 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5881 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5941 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6001 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6061 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6121 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6181 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6241 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6301 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6361 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6421 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6481 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6541 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6601 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6661 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6721 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6781 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6841 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6901 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6961 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7021 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7081 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7141 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7201 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7261 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7321 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7381 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7441 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7501 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7561 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7621 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7681 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7741 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7801 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     7861 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7921 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7981 agaatctgct tagg
//
